The Expression Of P3H4 In Hepatocellular Carcinoma And Its Correlation With Clinical Prognosis And Biological Function

Objective The aim of the present study was to explore the expression of P3H4 in hepatocellular carcinoma with its probably regulatory mechanisms and roles based on the HCC transcription dataset in The Cancer Genome Atlas(TCGA);to investigate the correlation between the expression of P3H4,clinicopathological factors and prognosis in HCC patients using clinical tissue samples.The effect of P3H4 on the biological function of hepatocellular carcinoma cells is initially discussed in order to provide reference for further research on its related mechanism.Methods First of all,this study analyzed the expression of P3H4 in human tumors and its relationship with the prognosis of HCC patients through multiple online analysis websites based on TCGA data and databases such as ENCORI,a prediction of P3H4 upstream ncRNAs regulate axis and its possible involvement in cellular functions and signaling pathways was done.Then,clinical samples were validated,q RT-PCR was used to detect the expression levels of P3H4 mRNA in 41 pairs of HCC cancer tissues and adjacent normal tissues.Immunohistochemistry was used to detect the expression of P3H4 in 102 HCC cancer tissues and 67 adjacent normal tissues.In addition,the clinicopathological data and follow-up information of these 102 HCC patients were collected.Chi-square test was used to analyze the correlation between P3H4 expression and clinicopathological parameters in patients with HCC;Survival analysis of P3H4 expression levels in HCC patients and subgroup was performed by Kaplan-Meier method,and comparisons between groups were performed by log-rank test.Cox proportional risk regression model was used to evaluate the prognostic risk factors.Next,selected P3H4 interfering lentivirus were used to construct two models of P3H4 down-regulation in Hep-3B and Huh-7 hepatocellular carcinoma cells,the effect of P3H4 interference was verified by q RT-PCR and Western blot.Finally,CCK-8 assay,Transwell assay,cell scratch assay and flow cytometry were respectively performed on the two hepatocellular carcinoma cell models,the effect of P3H4 on the proliferation,migration,invasion and apoptosis of hepatocellular carcinoma cells was analyzed.Results 1.Bioinformatics analysis showed that:(1)The expression of P3H4 in cancer tissues was up-regulated in bladder urothelial carcinoma(BLCA),breast invasive carcinoma(BRCA),cholangiocarcinoma(CHOL),esophageal carcinoma(ESCA),head and neck squamous cell carcinoma(HNSC),Renal papillary cell carcinoma(KIRP),Liver hepatocellular carcinoma(LIHC),lung adenocarcinoma(LUAD),lung squamous cell carcinoma(LUSC)and gastric adenocarcinoma(STAD).The only down-regulated expression was in renal chromophobe carcinoma(KICH);(2)The overall survival of HCC patients with high P3H4 expression was significantly shorter compared to that with low/moderate expression of P3H4;(3)It was predicted that miR-29c-3p may be the upstream targeted binding of P3H4 in hepatocellular carcinoma,three lncRNAs(DUXAP8,LINC00511 and CRNDE)may act as competitive regulatory ceRNAs;(4)The expression of P3H4 in HCC was significantly positively correlated with six types of immune infiltrating cells(B cells,CD8+T cells,CD4+T cells,macrophages,neutrophils and dendritic cells);(5)The expression of P3H4 in HCC was significantly positively correlated with the expressions of PDCD1,CD274 and CTLA-4;(6)P3H4 co-expression gene sets in HCC were mainly enriched in the regulation of cell growth and differentiation:such as protein binding,kinase activation,intracellular signal transduction,as well as viral oncogenic pathways,Fox O signaling pathways,central carbon in cancer Metabolism,VEGF signaling pathway and other cancer-related pathways.2.Clinical samples and cell function experiments showed that:(1)P3H4mRNA and protein were expressed in both HCC cancer tissues and adjacent normal tissues,but their expression levels in cancer tissues were significantly higher than that in adjacent normal tissues;(2)P3H4 protein expression level in HCC was significantly correlated with tumor size,MVI and BCLC stage;(3)HCC patients with high P3H4 expression had a worse prognosis than those with low expression,and P3H4 was an independent risk factor for OS and DFS in HCC patients.(4)Subgroup analysis showed that P3H4 was significantly correlated with OS and DFS in HCC patients without microvascular invasion;(5)Down-regulation of P3H4 expression in Hep-3B and Huh-7 HCC cells can inhibited cell proliferation,migration,invasion and promote cell apoptosis.Conclusion P3H4 was significantly highly expressed in HCC,and its expression level was closely associated with multiple malignant clinicopathological features as well as poor prognosis.P3H4 was an independent risk factor for the prognosis of HCC patients and a predictor of the prognosis of HCC patients without microvascular invasion.Down-regulation of P3H4 expression can inhibit the proliferation,migration and invasion of HCC cells and promote cell apoptosis.In a word,P3H4 may be play an important role in the occurrence and development of HCC,it may serve as a promising predictor of the prognosis.